Multiplex reverse transcription-polymerase chain reaction combined with on-chip electrophoresis as a rapid screening tool for candidate gene sets

The increasing number of factors to be taken into account in the oestrogen transcriptional process has created a need to develop a rapid screening method to evaluate their role in physiology and pathology. Molecular biology techniques enable gene expression studies at the mRNA level with small amounts of tissues. We therefore developed a semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) technique using fluorescent oligonucleotides to analyse simultaneously a large panel of interrelated genes involved in the oestrogen transcriptional pathway using a moderately expressed housekeeping gene, the hypoxanthine phosphoribosyltransferase gene (HPRT), as the reference gene. Expression levels of oestrogen receptors (ERalpha, ERbeta), cofactors AIB1, RIP140, SMRT and the Fas-associated protein-tyrosine phophatase-1 (FAP-1) genes were evaluated in breast, endometrial and ovarian cancer cell lines and in three ERalpha-positive and three ERalpha-negative breast cancer tumours. This technique provides a rapid and reliable way to quantitate simultaneously numerous mRNAs of genes involved in the oestrogen pathway from small amounts of tissues.

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Candidate gene biodosimeters of mice and human exposure to ionizing radiation by quantitative reverse transcription polymerase chain reaction

Journal of Cancer Research and Therapeutics ◽

10.4103/0973-1482.160912 ◽

2015 ◽

Vol 11 (3) ◽

pp. 549 ◽

Cited By ~ 5

Author(s):

Alireza Shirazi ◽

Hamed Rezaeejam ◽

Majid Valizadeh ◽

Pantea Izadi

Keyword(s):

Polymerase Chain Reaction ◽

Ionizing Radiation ◽

Candidate Gene ◽

Reverse Transcription ◽

Human Exposure ◽

Chain Reaction ◽

Polymerase Chain

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Development of a reverse transcription (RT) polymerase chain reaction (PCR) method for the detection of human norovirus in bivalve molluscs

Water Science & Technology ◽

10.2166/wst.2021.048 ◽

2021 ◽

Author(s):

Manjusha Lekshmi ◽

Sanath H. Kumar ◽

Kooloth Valappil Rajendran ◽

Binaya Bhusan Nayak

Keyword(s):

Polymerase Chain Reaction ◽

Reverse Transcription ◽

Rapid Screening ◽

Bivalve Molluscs ◽

Chain Reaction ◽

Reverse Transcription Pcr ◽

Pcr Method ◽

Polymerase Chain ◽

Rna Polymerase Gene ◽

Single Set

Abstract Noroviruses are significant seafood-borne pathogens, commonly associated with the consumption of filter feeding bivalve molluscs. Here, we report the development of a reverse transcription polymerase chain reaction (RT-PCR) method using primers based on the RNA-dependent RNA polymerase gene of norovirus genogroup II (NoV GII). Samples of bivalves were processed for the concentration of virus and extraction of RNA, followed by reverse transcription PCR. A total of 50 molluscan shellfish samples were analyzed, of which 16 samples yielded positive amplifications of norovirus nucleic acid. The PCR method described here, involving a single set of primers, is useful for rapid screening of shellfish for NoV GII.

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